Abstract
Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes inflammation and destruction of the joints as well as an increased risk of cardiovascular disease. RA synovial fibroblasts (RASFs) are involved in the progression of RA and release pro-inflammatory cytokines. On the other hand, microRNAs (miRs) may help control the inflammatory response of immune and non-immune cells. Therefore, our study used lentiviral expression vectors to test the effects of miR-126 overexpression on RASF proliferation and apoptosis. Luciferase experiments verified the targeting relationship between miR-126 and PIK3R2 gene. The co-transfection of anti-miR-126 and PIK3R2 siRNA to RASFs were used to identify whether PIK3R2 was directly involved in proliferation and apoptosis of miR-126-induced RASFs. Real-time polymerase chain reaction (PCR) was used to detect miR-126 and PIK3R2 expressions. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis and cell cycle. Western blotting was used to detect PIK3R2, PI3K, AKT and p-AKT proteins. After Lv-miR-126 infected RASFs, the relative expression of miR-126 was significantly enhanced. MiR-126 promoted RASF proliferation and inhibited apoptosis. Levels of PIK3R2 decreased while total PI3K and p-AKT levels increased in RASFs overexpressing miR-126. Co-transfection of anti-miR-126 and PIK3R2 siRNA also increased PI3K and p-AKT levels as well as RASF proliferation and reduced apoptosis, as compared to anti-miR-126 treatment alone. Finally, luciferase reporter assays showed that miR-126 targeted PIK3R2. Our data indicate that miR-126 overexpression in RASFs inhibits PIK3R2 expression and promotes proliferation while inhibiting apoptosis. This suggests inhibiting miR-126 may yield therapeutic benefits in the treatment of RA.
Highlights
Rheumatoid arthritis (RA), as a chronic and autoimmune disease of the joints, affects approximately 0.5-1% of adults worldwide [1, 2]
The results showed that compared with the RA synovial fibroblasts (RASFs)-Lv-vector and blank groups, total PI3K and p-AKT levels were increased in the RASFs of RASFs-Lv-miR126 group (Figure 6C)
We aimed to explore the effects of miR-126 on the proliferation and apoptosis of RASFs
Summary
Rheumatoid arthritis (RA), as a chronic and autoimmune disease of the joints, affects approximately 0.5-1% of adults worldwide [1, 2]. About 30% of RA patients would become permanently work disabled in the first 2-3 years without sufficient treatment [3]. An improved understanding of RA pathogenesis has led to various modern therapeutic options; more research is needed to reduce side effects and improve the quality of life of patients [4]. RA synovial fibroblasts (RASFs) are major effectors of joint destruction and joint inflammation and play a critical role in the pathogenesis of RA, contributing to the formation of rheumatoid pannus [5]. RASFs can spontaneously secrete numerous pro-inflammatory cytokines, as well as innate immunity and matrixdegradation products, which promote RA pathogenesis [7]
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